Chris:LabNotes/FateMapping/Calendar/2014/2014-10-30: Difference between revisions
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*Follow online published protocol (make sure to prep using specifications for smaller D5020 kit) | *Follow online published protocol (make sure to prep using specifications for smaller D5020 kit) | ||
*Final elution = 10ul | *Final elution = 10ul | ||
*'''Note: Can quantify bisulfite conversion recovery by using Qubit ssDNA assay''' |
Revision as of 22:03, 5 November 2014
Bisulfite Conversion Test 3
Background
- The previous two tests didn't seem to have worked (CircLigase didn't seem to have circularized either the sample or the positive control)
- This time, I'll repeat the experiment but only use CircLigase II
Procedure
- Below are the reactions I want to do for this experiment:
Rxn 1 (gDNA) Bisulfite Conversion Rxn 2 (gDNA) Bisulfite Conversion -> T4 PNK -> CircLigase II -> Exo I/III Rxn 3 (NTC) Bisulfite Conversion Rxn 4 (NTC) Bisulfite Conversion -> T4 PNK -> CircLigase II -> Exo I/III
EZ DNA Methlyation-Direct
- Use human gDNA (110ng/ul) for template for rxn's 1->4 (use 3.64ul to get 400ng of gDNA)
- Follow online published protocol (make sure to prep using specifications for smaller D5020 kit)
- Final elution = 10ul
- Note: Can quantify bisulfite conversion recovery by using Qubit ssDNA assay